Journal: Journal of Ovarian Research

Article Title: FTO attenuates the cytotoxicity of cisplatin in KGN granulosa cell-like tumour cells by regulating the Hippo/YAP1 signalling pathway

doi: 10.1186/s13048-024-01385-5

Figure Lengend Snippet: Cisplatin promoted the KGN cells apoptosis, decreased the expression of FTO, and activated the Hippo/YAP signaling pathway, in a dose dependent manner. ( A-B ) Flow cytometry assays were performed to determine the effect of cisplatin on KGN cells apoptosis. And a quantitative analysis of the rate of apoptosis was performed. ( C-M ) Western blot assay was applied to detect the effect of cisplatin on the protein expression level of apoptosis related protein, including Bcl-2, BAX, and cleaved-caspase-3, FTO and the key molecular of Hippo/YAP signaling pathway. β-actin was used as a loading control. ( N-O ) Immunofluorescence assay was performed to detect the effect of cisplatin on the expression and location of YAP1. (scale bar: 100 μm). Student’s t-test. The results were showed mean ± SEM from three independent experiments. (α = 0.05, * P < 0.05, ** P < 0.01, *** P < 0.001)

Article Snippet: Blocked the cells in 5% bovine serum albumin (BSA) for 1 h and incubated the cells with rabbit monoclonal antibody against YAP1 (1:200, Proteintech) at 4 °C overnight.

Techniques: Expressing, Flow Cytometry, Western Blot, Control, Immunofluorescence

Journal: Journal of Ovarian Research

Article Title: FTO attenuates the cytotoxicity of cisplatin in KGN granulosa cell-like tumour cells by regulating the Hippo/YAP1 signalling pathway

doi: 10.1186/s13048-024-01385-5

Figure Lengend Snippet: Overexpression of FTO decreased KGN cells apoptosis and disrupted the Hippo/YAP signaling pathway. ( A ) The expression of FTO, Bc-2, BAX, MST, p-MST, LATS1, p-LATS1, YAP1, and p-YAP1 were monitored by western blotting. β-actin was used as a loading control. ( B-K ) Quantitative comparison of expression levels of FTO, Bcl-2, BAX, p-MST1/MST1, p-LATS1/LATS1, p-YAP1/YAP1 in different groups. ( L-N ) The mRNA expression level of CTGF, CYR61, and ANKRD1 were determined by qRT-PCR. GAPDH was used as a loading control. Student’s t-test. The results were showed mean ± SEM from three independent experiments. (α = 0.05, * P < 0.05, ** P < 0.01, *** P < 0.001). (O - P) Immunofluorescence assay was performed to determine the effect of FTO on the expression and location of YAP1. (scale bar: 100 μm)

Article Snippet: Blocked the cells in 5% bovine serum albumin (BSA) for 1 h and incubated the cells with rabbit monoclonal antibody against YAP1 (1:200, Proteintech) at 4 °C overnight.

Techniques: Over Expression, Expressing, Western Blot, Control, Comparison, Quantitative RT-PCR, Immunofluorescence

Journal: Journal of Ovarian Research

Article Title: FTO attenuates the cytotoxicity of cisplatin in KGN granulosa cell-like tumour cells by regulating the Hippo/YAP1 signalling pathway

doi: 10.1186/s13048-024-01385-5

Figure Lengend Snippet: Downregulation of FTO promoted KGN cells apoptosis and activated the Hippo/YAP signaling pathway. ( A ) Western blotting was used to detect the effects of si-FTO on the expression of FTO, Bcl − 2, BAX, MST, p-MST, LATS, p-LATS1, YAP1, and p - YAP1. β-actin was used as a loading control. ( B-K ) Quantitative comparison of expression levels of FTO, Bcl-2, BAX, p-MST1/MST1, p-LATS1/LATS1, p-YAP1/YAP1 in different groups. ( L-N ) The mRNA expression level of CTGF, CYR61, and ANKRD1 were determined by qRT-PCR. GAPDH was used as a loading control. Student’s t-test. The results were showed mean ± SEM from three independent experiments. (α = 0.05, * P < 0.05, ** P < 0.01, *** P < 0.001). ( O-P ) Immunofluorescence assay was performed to determine the effect of si-FTO on the expression and location of YAP1. (scale bar: 100 μm)

Article Snippet: Blocked the cells in 5% bovine serum albumin (BSA) for 1 h and incubated the cells with rabbit monoclonal antibody against YAP1 (1:200, Proteintech) at 4 °C overnight.

Techniques: Western Blot, Expressing, Control, Comparison, Quantitative RT-PCR, Immunofluorescence

Journal: Journal of Ovarian Research

Article Title: FTO attenuates the cytotoxicity of cisplatin in KGN granulosa cell-like tumour cells by regulating the Hippo/YAP1 signalling pathway

doi: 10.1186/s13048-024-01385-5

Figure Lengend Snippet: Overexpression of FTO promoted cisplatin-induced injured KGN cells proliferation and decreased their apoptosis via Hippo/YAP signaling pathway, while VP reversed it. ( A ) Western blotting was used to detect the effects of FTO on the expression of Bcl-2, BAX, cleaved-caspase-3, PCNA, MST, p-MST, LATS1, p-LATS1, YAP1, and p-YAP1in injured KGN cells. β-actin was used as a loading control. ( B-K ) Quantitative comparison of expression levels Bcl-2, BAX, cleaved-caspase-3, PCNA, MST, p-MST, LATS1, p-LATS1, YAP1, and p-YAP1 in different groups. ( L ) CCK-8 assay was performed to measure the rate the KGN cells with different treatments. ( M-N ) Flow cytometry assays were performed to determine the apoptosis rate in different groups, and the quantitative comparison was included. One-way ANOVA followed by Tukey’s multiple comparison test. The results were showed mean ± SEM from 3 independent experiments. (α = 0.05, * P < 0.05, ** P < 0.01, *** P < 0.001). ( O-P ) Immunofluorescence assay was performed to determine the expression and location of YAP1 in different groups. (scale bar: 100 μm)

Article Snippet: Blocked the cells in 5% bovine serum albumin (BSA) for 1 h and incubated the cells with rabbit monoclonal antibody against YAP1 (1:200, Proteintech) at 4 °C overnight.

Techniques: Over Expression, Western Blot, Expressing, Control, Comparison, CCK-8 Assay, Flow Cytometry, Immunofluorescence

Journal: Journal of Ovarian Research

Article Title: FTO attenuates the cytotoxicity of cisplatin in KGN granulosa cell-like tumour cells by regulating the Hippo/YAP1 signalling pathway

doi: 10.1186/s13048-024-01385-5

Figure Lengend Snippet: Downregulation of FTO promoted the cisplatin-induced KGN cells injury, and VP could aggregate it. ( A ) Western blotting was used to detect the effects of si -FTO on the expression of Bcl-2, BAX, cleaved-caspase-3, PCNA, MST, p-MST, LATS1, p-LATS1, YAP1, and p-YAP1in injured KGN cells. β-actin was used as a loading control. ( B-K ) Quantitative comparison of expression levels Bcl-2, BAX, cleaved-caspase-3, PCNA, MST, p-MST, LATS1, p-LATS1, YAP1, and p-YAP1 in different groups. ( L ) CCK-8 assay was performed to measure the rate the KGN cells with different treatments. ( M-N ) Flow cytometry assays were performed to determine the apoptosis rate in different groups, and the quantitative comparison was included. One-way ANOVA followed by Tukey’s multiple comparison test. The results were showed mean ± SEM from 3 independent experiments. (α = 0.05, * P < 0.05, ** P < 0.01, *** P < 0.001). ( O-P ) Immunofluorescence assay was performed to determine the expression and location of YAP1 in different groups. (scale bar: 100 μm)

Article Snippet: Blocked the cells in 5% bovine serum albumin (BSA) for 1 h and incubated the cells with rabbit monoclonal antibody against YAP1 (1:200, Proteintech) at 4 °C overnight.

Techniques: Western Blot, Expressing, Control, Comparison, CCK-8 Assay, Flow Cytometry, Immunofluorescence